Development and optimization of a sensitive pseudovirus-based assay for HIV-1 neutralizing antibodies detection using A3R5 cells
Sensitive assays for HIV-1 neutralizing antibody detection are urgently needed for vaccine immunogen optimization and identification of protective immune response levels. In this study, we developed an easy-to-use HIV-1 pseudovirus neutralization assay based on a human CD4+ lymphoblastoid cell line...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Taylor & Francis Group
2018-01-01
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| Series: | Human Vaccines & Immunotherapeutics |
| Subjects: | |
| Online Access: | http://dx.doi.org/10.1080/21645515.2017.1373922 |
| _version_ | 1797677534528143360 |
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| author | Qingqing Chen Jianhui Nie Weijin Huang Yanmei Jiao Lan Li Tong Zhang Juan Zhao Hao Wu Youchun Wang |
| author_facet | Qingqing Chen Jianhui Nie Weijin Huang Yanmei Jiao Lan Li Tong Zhang Juan Zhao Hao Wu Youchun Wang |
| author_sort | Qingqing Chen |
| collection | DOAJ |
| description | Sensitive assays for HIV-1 neutralizing antibody detection are urgently needed for vaccine immunogen optimization and identification of protective immune response levels. In this study, we developed an easy-to-use HIV-1 pseudovirus neutralization assay based on a human CD4+ lymphoblastoid cell line A3R5 by employing a high efficient pseudovirus production system. Optimal conditions for cell counts, infection time, virus dose and concentration of DEAE-dextran were tested and identified. For T-cell line-adapted tier 1 virus strains, significantly higher inhibitory efficiency was observed for both monoclonal neutralizing antibody (4 fold) and plasma (2 fold) samples in A3R5 than those in TZM-bl assay. For circulating tier 2 strains, the A3R5 pseudovirus assay showed even much higher sensitivity for both neutralizing antibody (10 fold) and plasma (9 fold) samples. When sequential neutralizing antibody seroconverting samples were tested in both A3R5 and TZM-bl assays, the seroconverting points could be detected earlier for tier 1 (15.7 weeks) and tier 2 (68.3 weeks) strains in A3R5 assay respectively. The high sensitive pseudovirus assay using more physiological target cells could serve as an alternative to the TZM-bl assay for evaluation of vaccine-induced neutralizing antibodies and identification of the correlates of protection. |
| first_indexed | 2024-03-11T22:46:34Z |
| format | Article |
| id | doaj.art-358cf4bab523415084b423a2538f4e33 |
| institution | Directory Open Access Journal |
| issn | 2164-5515 2164-554X |
| language | English |
| last_indexed | 2024-03-11T22:46:34Z |
| publishDate | 2018-01-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | Human Vaccines & Immunotherapeutics |
| spelling | doaj.art-358cf4bab523415084b423a2538f4e332023-09-22T08:17:52ZengTaylor & Francis GroupHuman Vaccines & Immunotherapeutics2164-55152164-554X2018-01-0114119920810.1080/21645515.2017.13739221373922Development and optimization of a sensitive pseudovirus-based assay for HIV-1 neutralizing antibodies detection using A3R5 cellsQingqing Chen0Jianhui Nie1Weijin Huang2Yanmei Jiao3Lan Li4Tong Zhang5Juan Zhao6Hao Wu7Youchun Wang8National Institutes for Food and Drug Control (NIFDC)National Institutes for Food and Drug Control (NIFDC)National Institutes for Food and Drug Control (NIFDC)Beijing You'an Hospital, Capital Medical UniversityBeijing You'an Hospital, Capital Medical UniversityBeijing You'an Hospital, Capital Medical UniversityNational Institutes for Food and Drug Control (NIFDC)Beijing You'an Hospital, Capital Medical UniversityNational Institutes for Food and Drug Control (NIFDC)Sensitive assays for HIV-1 neutralizing antibody detection are urgently needed for vaccine immunogen optimization and identification of protective immune response levels. In this study, we developed an easy-to-use HIV-1 pseudovirus neutralization assay based on a human CD4+ lymphoblastoid cell line A3R5 by employing a high efficient pseudovirus production system. Optimal conditions for cell counts, infection time, virus dose and concentration of DEAE-dextran were tested and identified. For T-cell line-adapted tier 1 virus strains, significantly higher inhibitory efficiency was observed for both monoclonal neutralizing antibody (4 fold) and plasma (2 fold) samples in A3R5 than those in TZM-bl assay. For circulating tier 2 strains, the A3R5 pseudovirus assay showed even much higher sensitivity for both neutralizing antibody (10 fold) and plasma (9 fold) samples. When sequential neutralizing antibody seroconverting samples were tested in both A3R5 and TZM-bl assays, the seroconverting points could be detected earlier for tier 1 (15.7 weeks) and tier 2 (68.3 weeks) strains in A3R5 assay respectively. The high sensitive pseudovirus assay using more physiological target cells could serve as an alternative to the TZM-bl assay for evaluation of vaccine-induced neutralizing antibodies and identification of the correlates of protection.http://dx.doi.org/10.1080/21645515.2017.1373922hiv-1neutralizing antibodypseudovirusa3r5tzm-bl |
| spellingShingle | Qingqing Chen Jianhui Nie Weijin Huang Yanmei Jiao Lan Li Tong Zhang Juan Zhao Hao Wu Youchun Wang Development and optimization of a sensitive pseudovirus-based assay for HIV-1 neutralizing antibodies detection using A3R5 cells Human Vaccines & Immunotherapeutics hiv-1 neutralizing antibody pseudovirus a3r5 tzm-bl |
| title | Development and optimization of a sensitive pseudovirus-based assay for HIV-1 neutralizing antibodies detection using A3R5 cells |
| title_full | Development and optimization of a sensitive pseudovirus-based assay for HIV-1 neutralizing antibodies detection using A3R5 cells |
| title_fullStr | Development and optimization of a sensitive pseudovirus-based assay for HIV-1 neutralizing antibodies detection using A3R5 cells |
| title_full_unstemmed | Development and optimization of a sensitive pseudovirus-based assay for HIV-1 neutralizing antibodies detection using A3R5 cells |
| title_short | Development and optimization of a sensitive pseudovirus-based assay for HIV-1 neutralizing antibodies detection using A3R5 cells |
| title_sort | development and optimization of a sensitive pseudovirus based assay for hiv 1 neutralizing antibodies detection using a3r5 cells |
| topic | hiv-1 neutralizing antibody pseudovirus a3r5 tzm-bl |
| url | http://dx.doi.org/10.1080/21645515.2017.1373922 |
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